Figure 2

Biofilm structure visualized by CLSM and TEM. CLSM images of a 64.5 h 9-species biofilm stained by multiplex FISH for (A) V. dispar (purple; VEI217-ROX, 40% formamide), C. rectus (blue; Camp655-Cy5, 30% formamide), F. nucleatum (red; Fnuc133c-Cy3, 30% formamide), and P. intermedia (green; L-Pint649-2-FAM, 30% formamide), (B) V. dispar (purple; VEI217-ROX, 40% formamide), A. naeslundii (red; L-Act476-2-Cy3, 25% formamide), S. intermedius (green; L-Sco/int172-2-FAM, 25% formamide), and S. oralis (blue; MIT447-Cy5, 25% formamide), and (C) V. dispar (purple; VEI217-ROX, 40% formamide), T. forsythia (green; Tfor582-FAM, 40% formamide), P. gingivalis (red; L-Pgin1006-Cy3, 30% formamide), and C. rectus (blue; Camp655-Cy5, 30% formamide). Images are 1-μm-thick transverse (large images), sagittal (right) and coronal (bottom) slices at the positions indicated by the fine lines. The length of the bars indicates 20 μm. (D) TEM image of a 64.5 h multi-species biofilm demonstrating the predominance of varius cocci or very short rods (S. oralis, S. intermedius, V. dispar, P. intermedia) and of the fusiform F. nucleatum cells. Bar = 5 μm.